Fertilization Process Research Articles

The Criminal Liability in the Context of Fertilizer Trade in Indonesia

Indonesia is an agricultural country and its agricultural sector which important in meeting food needs. On its journey, the agricultural sector in Indonesia is developing to improve the quality and production of food needs. Then, the fertilizer process has an important role in increasing the production of agricultural products, and the government needs to implement policies to subsidize the price of fertilizers for fertilizer distributors and arrange the regulations underlying the policy. If the fertilizer trade for fertilizer distributors is based on more than clear regulations, how can Indonesia's agricultural journey run. Thus, this study aims to attempt to describe the criminal liability of fertilizer trade based on court decisions at the first level up to the cassation level (namely, Decision Number. 137/pid.sus/2020/PN.Btg, Court of Appeal (Number. 487/Pid.Sus/2020/PT SMG and Cassation Decision Number. 78/Pid.Sus/2022). This study is normative research with a case approach, requiring secondary data in legal documents and references relevant to the subject matter. Data were collected by identification and inventory and then analyzed using the basic principles of criminal law. The novelty is fulfilling criminal liability is not enough regarding the psychological aspects (adult, healthy mind). However, it must also fulfill the normative measure of the act. The results showed that the defendant in the first instance and appellate level decisions were able to take liabilitybut was not sentenced because the element of the action in the prosecutor's charge, which stated "conducting trading business activities without having a license in the field of trade granted by the Ministry" was not fulfilled/proven, because the defendant had a trading business license (SIUP) issued by the Integrated Investment and Licensing Agency Number: 519/054/2014 dated June 04, 2014, under the name of the Agricultural Facilities and Organic Fertilizer Shop "Tani Jaya." The cassation verdict of the defendant can be held responsible and sentenced to punishment because measures from the psychological aspect are fulfilled. The measurements are from the normative aspect demanded by the public prosecutor, especially the elements that are also fulfilled. It can be consluded that based on the court's decision to measure the perpetrator's liability for a criminal offense, two measures must be met in imposing punishment.

Ribonuclease Inhibitor 1 (RNH1) Regulates Sperm tsRNA Generation for Paternal Inheritance through Interacting with Angiogenin in the Caput Epididymis.

Environmental stressors can induce paternal epigenetic modifications that are a key determinant of the intergenerational inheritance of acquired phenotypes in mammals. Some of them can affect phenotypic expression through inducing changes in tRNA-derived small RNAs (tsRNAs), which modify paternal epigenetic regulation in sperm. However, it is unclear how these stressors can affect changes in the expression levels of tsRNAs and their related endonucleases in the male reproductive organs. We found that Ribonuclease inhibitor 1 (RNH1), an oxidation responder, interacts with ANG to regulate sperm tsRNA generation in the mouse caput epididymis. On the other hand, inflammation and oxidative stress induced by either lipopolysaccharide (LPS) or palmitate (PA) treatments weakened the RNH1-ANG interaction in the epididymal epithelial cells (EEC). Accordingly, ANG translocation increased from the nucleus to the cytoplasm, which led to ANG upregulation and increases in cytoplasmic tsRNA expression levels. In conclusion, as an antioxidant, RNH1 regulates tsRNA generation through targeting ANG in the mouse caput epididymis. Moreover, the tsRNA is an epigenetic factor in sperm that modulates paternal inheritance in offspring via the fertilization process.

Heat stress mediates toxicity of rutile titanium dioxide nanoparticles on fertilisation capacity in the broadcast spawning mussel Mytilus galloprovincialis

Titanium dioxide nanoparticle (nTiO2) pollution of marine environments is rapidly increasing with potentially deleterious effects on wildlife. Yet, the impacts of nTiO2 on reproduction remain poorly understood. This is especially the case for broadcast spawners, who are likely to be more severely impacted by environmental disturbances because their gametes are directly exposed to the environment during fertilisation. In addition, it is unclear whether rising water temperatures will further exacerbate the impact of nTiO2 toxicity. Here, in a series of fertilisation trials, we systematically examine the main and interactive effects of nTiO2 exposure and seawater temperature on fertilisation success in the Mediterranean mussel Mytilus galloprovincialis. Specifically, our fertilisation trials explored whether nTiO2 exposure influences fertilisation rates when (i) eggs alone are exposed, (ii) both sperm and eggs are exposed simultaneously, and (iii) whether increases in seawater temperature interact with nTiO2 exposure to influence fertilisation rates. We also ask whether changes in nTiO2 concentrations influence key sperm motility traits using computer-assisted sperm analysis (CASA). In fertilisation trials for treatment groups (i) and (ii), we found no main effects of nTiO2 at environmentally relevant concentrations of 5, 10 and 50 μg L−1 on fertilisation capacity relative to the control. Consistent with these findings, we found no effect of nTiO2 exposure on sperm motility. However, in treatment group (iii), when fertilisation trials were conducted at higher temperatures (+6 °C), exposure of gametes from both sexes to 10 μg L−1 nTiO2 led to a reduction in fertilisation rates that was significantly greater than when gametes were exposed to elevated temperature alone. These interacting effects of nTiO2 exposure and seawater temperature demonstrate the toxic potential of nTiO2 for fertilisation processes in a system that is likely to be impacted heavily by predicted future increases in sea surface temperatures.

Analysis of Maize Planting Mode and Simulation and Optimization of Ridging and Fertilization Components in Arid Area of Northwest China

The arid area of Northwest China belongs to the rain-fed agricultural area of the Loess Plateau, and water resources have become one of the important factors limiting agricultural development in this area. This study employed the AquaCrop model to predict the yield advantages and environmental adaptability of maize in Dingxi City from 2016 to 2020 under two cultivation practices: ridge tillage (100% film coverage with double ridge-furrow planting) and flat planting (81.8% film coverage with wide-film planting). The numerical simulation of the tillage and fertilization process of the double-ridge seedbed was carried out by EDEM, and the key components were tested by the Box–Behnken center combination test design principle to obtain the optimal parameter combination. The results showed that ridge planting was more suitable for agricultural planting in rain-fed arid areas in Northwest China. The simulation analysis of ridging and fertilization showed that the forward speed of the combined machine was 0.50 m/s, the rotation speed of the trough wheel of the fertilizer discharger was 39 rmp, and the rotary tillage depth was 150 mm. The qualified rate of seedbed tillage was 93.6%, and the qualified rate of fertilization was 92.1%. The research shows that the whole-film double ridge-furrow sowing technology of maize is more suitable for the rain-fed agricultural area in the arid area of Northwest China. The simulation results of the ridging fertilization device are consistent with the field experiment results. The research results provide a certain technical reference for the optimization of the whole-film double ridge-furrow sowing technology.

Cooler Is Better: The Role of Temperature in the Reproductive Biology of Persian Walnut (Juglans regia L.) Studied in a Hungarian-Bred Cultivar

The aim was to determine the growth of pollen tubes throughout the transmitting tissue and the process of fertilization in open, optimal and over pollination on the 3rd, 6th and 9th day after pollination of pistillate flowers, together with apomixis. The experiment was carried out over 2 years in Lovasberény, Fejér district, Hungary, which differed in temperature conditions. The average number of pollen tubes in the locule of the ovary was significantly higher in 2017 (1.3%) than in 2019 (1%). A higher percentage of fruit set was recorded in 2017 compared to 2019. The percentage of apomictic fruits from variant of unpollination was higher in 2017 (10%) than in 2019 (3.7%). Based on these results it can be concluded that pollen tube growth and post-fertilization processes together with apomixis of cultivar ‘AK’ are better adaptable to lower temperatures. These findings may help us to move walnut production to the north or to higher altitudes, where the climate is more continental.

Techno-economic assessment of industrial food waste composting facility: Evaluating bulking agents, processing strategies, and market dynamics

Techno-economic assessment (TEA) of a valorization of bulking agent (BA) ratios on the food waste compost value chain is made to assess economic feasibility. TEA was performed with two plans (Plan A: existing composting facilities; Plan B: new composting facilities) and each plan was under four scenarios. The BA (i.e. corn stalks, garden waste, and watermelon seedlings) ratio of 5 % (S1), 10 % (S2), 20 % (S3), and garden waste with a ratio of 20 % (S4). Results indicate that S2, with a net present value (NPV) of 128.9 million, represents Plan A’s most economically viable scenario. Although the total operating costs of S4 were 18.9 %–23.5 % higher, 25.6 %–42.2 % higher total revenue made S4 have an NPV of 92.9 million, making it the most viable scenario in Plan B. All scenarios show positive NPV within a ± 20 % fluctuation range. Organic fertilizer price, government subsidies, and processing capacity were the key factors influencing NPV.

Negotiating Work and Family Spheres: The Dyadic Effects of Flexible Work Arrangements on Fertility Among Dual-Earner Heterosexual Couples

Academics and policymakers have suggested making flexible work arrangements (FWAs) the default in workplaces to promote a family-friendly workplace culture conducive to having and raising children. However, systematic research investigating how FWAs, as a long-term approach to negotiating work–family spheres, are related to fertility among dual-earner heterosexual couples is limited. Drawing on the linked-lives perspective, we theorize the relationship between FWAs and fertility among couples and potential variation depending on the interplay of both spouses’ work and family characteristics. We test our hypotheses using longitudinal couple-level dyadic data in the United Kingdom (2010–2022). We find that although FWA availability alone is unrelated to fertility, wives’ (not husbands’) FWA use is significantly associated with a higher probability of experiencing a first birth. Moreover, the effect of wives’ FWA use is particularly pronounced when both spouses work in professional and managerial occupations and when husbands contribute a larger proportion of income and at least equal housework. This study reveals a gendered effect of FWAs on fertility across work–family arrangements, deepening our understanding of couple-level dynamics in the fertility process.

Reproduction and Follow-up of Embryonic Development of Achatina fulica

Although Achatina fulica is hermaphrodite, the fertilization process in this type of snail occurs internally and in conjunction, and the mechanism of reproduction is to place the eggs in groups in a hole and then cover them with dirt. The stages of embryonic development from laying the eggs until the hatching process took ten days and after hatching. A small individual similar to the parents emerges. The criterion for embryonic development in this study focused on the noticeable increase in the size of the fetus with the progression of time or the age of the fetus during the egg incubation period.

Environmental Performance of Ammonia Production in Indonesia using Life Cycle Assessment Approach

Ammonia is a fundamental component in fertilizer and chemical manufacturing processes around the world, but its production is a significant contributor to CO2 emissions in chemical industry. Implementation of carbon capture utilization and storage (CCUS) offers an alternative decarbonisation strategy to mitigate CO2 emissions in ammonia production. This study assesses the environmental performance of ammonia production through life cycle assessment (LCA) methodology. Environmental impacts are calculated using openLCA software with various impact assessment methods, including CML IA Baseline, Impact 2002+, Recipe 2016 Midpoint (H), and AWARE. The study scope encompasses the cradle-to-grave analysis, from the extraction of raw materials and transportation to ammonia production, main processes, distribution, and consumer product consumption, with a declared unit of 1-kg ammonia product. Our findings showed that CO2 removal and Power Plant in core processes in the core process as the most significant contributors to Global Warming Potential. Therefore, sensitivity analysis was conducted by reducing CO2 emission by 10% and 70% through CCUS implementation. The results showed that the CCUS implementation could reduce Global Warming Potential by up to 43%.

Climate Change and Soil Fertility: A Review of Strategies for a Sustainable Food Security in India

The long-term food security of humanity on Earth will be significantly influenced by the effects of climate change soils and then the agricultural production. In addition to being vulnerable to climate change, agriculture also contributes significantly to it. The article explores how soil fertility processes and qualities are impacted by climate change on a worldwide scale and adaptation and mitigation options for a sustainable food security. Due to the intricate relationship between soils and the climate system through nutrient and hydrologic cycles, it is anticipated that changes in the global climate may have an effect on soil fertility through changes in the physical, chemical, and biological characteristics of the soil as a result of rising temperatures, altered precipitation patterns, increased atmospheric concentrations of greenhouse gases, and other factors.

A bovine oviduct microfluidic platform to evaluate the effects of cisplatin and hormones on cilia beating frequencies

Chemotherapeutics such as cisplatin modulate the function of cells in fallopian tubes (oviducts) reducing the formation of sperm reservoirs that promote gamete interaction during the fertilization process. The mechanisms behind these resulting deficiencies must be better understood to improve fertility for cancer patients. A microfluidic platform was developed by immobilizing a transparent polycarbonate membrane between two poly(dimethyl siloxane) molds to make channels for the growth of a functional monolayer of oviduct cells on one side and medium on the other side. Freshly isolated bovine oviduct epithelial cells were cultured on the membrane with an air-liquid interface after 5 days, and the resulting cells had consistent cilia beating frequencies (CBF) for 12 weeks between 4.92 ± 0.59 Hz and 5.76 ± 0.94 Hz. Incubation with 75 pg ml−1 17-beta-estradiol and 100 ng ml−1 progesterone increased the CBF from 4.62 ± 0.16 Hz to 7.61 ± 0.41 Hz. When incubated with 50 µM of cisplatin, the CBF dropped by 40 ± 19 % without hormones and 52 ± 9 % with hormones, with similarly decreased CBFs. Short 2-h incubations of bovine oviduct cells with cisplatin directly reduced the beating of ciliated cells, illustrating that the short-term effects (hours) of chemotherapeutics should be considered in oncofertility.

Site-specific N-glycoproteomic analysis reveals up-regulated fucosylation in seminal plasma of asthenozoospermia.

N-linked glycoproteins are rich in seminal plasma, playing essential roles in supporting sperm function and fertilization process. The alteration of seminal plasma glycans and its correspond glycoproteins may lead to sperm dysfunction and even infertility. In present study, an integrative analysis of glycoproteomic and proteomic was performed to investigate the changes of site-specific glycans and glycoptoteins in seminal plasma of asthenozoospermia. By large scale profiling and quantifying 5,018 intact N-glycopeptides in seminal plasma, we identified 92 intact N-glycopeptides from 34 glycoproteins changed in asthenozoospermia. Especially, fucosylated glycans containing lewis x, lewis y and core fucosylation were significantly up-regulated in asthenozoospermia compared to healthy donors. The up-regulation of fucosylated glycans in seminal plasma may interfere sperm surface compositions and regulation of immune response, which subsequently disrupts sperm function. Three differentiated expression of seminal vesicle-specific glycoproteins (fibronectin, seminogelin-2, and glycodelin) were also detected with fucosylation alteration in seminal plasma of asthenozoospermia. The interpretation of the altered site-specific glycan structures provides data for the diagnosis and etiology analysis of male infertility, as well as providing new insights into the potential therapeutic targets for male infertility.

Impact of fluazuron on oocyte maturation: May the antiparasitic affect bovine reproduction?

Fluazuron is a novel veterinary pour-on antitick formulation which can be applied simultaneously with bovine reproduction management strategies. Considering the economic importance of the livestock industry in many countries, it is important to know whether antiparasitics such as fluazuron may cause embryonic loss. The aim of this study was to evaluate the toxicological effect of fluazuron on bovine oocytes during in vitro maturation. The best fluazuron concentrations were determined in a preliminary experiment on Chinese hamster ovary (CHO)-K1 cells and further used to compare fluazuron toxicity in both study models. Results of the annexin V and alkaline single cell gel electrophoresis assays demonstrated that fluazuron caused cytotoxicity and genotoxicity in bovine cumulus cells at all the concentrations tested (50, 75 and 100 μg fluazuron/mL). The evaluation of cortical granules and mitochondria distribution showed that cytoplasmic maturation was not affected by fluazuron treatment. However, a decrease in metaphase II + polar body, degenerate oocytes as well as disorganized chromatin in polar body were observed at all concentrations tested. Whereas the fertilization process was not altered by 50 μg/mL fluazuron, the embryo development rate decreased significantly. No significant differences were observed in any of the oxidative stress parameters assessed. This study contributes to a better understanding of fluazuron in bovines, suggesting that the antiparasitic may affect bovine reproduction and might cause embryo loss.

Pelatihan Petani dalam Produksi Pupuk Organik di Desa Tegalgede, Kecamatan Sumbersari, Kabupaten Jember

Chemical fertilizers are increasingly rare and expensive, causing farmers (partners) to look for solutiosn by using organic fertilizer as a combination. However, a problem is currently being faced by partners, namely related to the production process of organic fertilizer made from goat manure. The characteristics of goat manure are hard, and it takes quite a long time to decompose, so farmers have to wait approximately one month before it can be applied as basic fertilizer on agricultural land. Efforts to speed up the decomposition process have been made by partners, namely by crushing or smoothing goat manure by pounding it manually. This stage requires quite a lot of time and energy, so it is felt to be less efficient. This is due to the lack of technology adopted in the partner environment. Based on the conditions and problems faced by partners, through this activity a solution will be offered in the form of technology for processing goat manure as raw material for organic fertilizer. This technology is in the form of a goat manure crushing machine to help speed up the decomposition process and provide nutrients, so that partners do not have to wait long for planting to start. This community service activity is carried out by providing counseling and training in making organic fertilizer from goat dung as raw material, accompanied by the use of a goat dung crushing machine. It is hoped that partners’ knowledge and skills can increase, especially in processing organic fertilizer made from goat manure.

The AutoSpawner system - Automated ex situ spawning and fertilisation of corals for reef restoration

The restoration of reefs damaged by global and local pressures remains constrained by the scale of intervention currently feasible. Traditional methods for ex situ sexual propagation of corals produce limited materials, typically of limited genetic diversity and only sufficient for small field trials. The development and validation of new technologies to upscale and automate coral propagation is required to achieve logistically and financially feasible reef restoration at ecologically relevant scales. To address the need for upscaled production of genetically diverse material for use in reef restoration we designed an automated system (the AutoSpawner) for harvesting, fertilising and washing gametes from tropical broadcast-spawning corals. The system includes a novel high density dynamic fertilisation process, which enables the production of large numbers of fertilised coral eggs (>7 million per night for highly fecund species) without any downstream negative effects on larval quality. The functionality of the system and the quality of the produced larvae was assessed using multiple species from two coral families (Acroporidae and Merulinidae) across a range of spawning and gamete characteristics. We present the schematics and protocols required for automated sexual propagation of high-quality coral larvae using this novel system; and demonstrate that the time demands, and labour costs, associated with traditional manual-based sexual propagation of corals can be reduced by up to 113-fold using the AutoSpawner.

P-014 Extracellular vesicles (EVs) from female reproductive tracts positively affect sperm capacitation and progressive motility

Abstract Study question Do EVs from human follicular fluid (FF-EVs), cervicovaginal fluid (CVF-EVs), and endometrial stromal cells spent medium (ESCs-EVs) induce sperm capacitation and enhance sperm motility? Summary answer In humans, EVs from different female reproductive tracts actively contribute to sperm maturation by inducing capacitation and enhancing sperm progressive motility. What is known already In the human body, EVs represent crucial players in cell-to-cell communication. EVs presence has been detected also in the female and male reproductive systems, specifically in seminal, follicular, oviductal, endometrial and vaginal fluid, as well as in embryo secretions. Moreover, EVs have been shown to be of pivotal importance in the fertilization process, as they support the maturation of spermatozoa and oocytes, as well as embryos in their development and endometrial implantation. The majority of these studies, though, have been conducted in animal models, underscoring the necessity to replicate analogous investigations also in humans. Study design, size, duration Follicular fluid (FF, n = 8), cervicovaginal fluid (CVF, n = 20) and endometrial tissue (n = 8) samples were collected from women undergoing assisted reproductive technology (ART) cycles at San Raffaele Hospital Fertility Centre (Milan). CVF were collected 2 and 7 days after the LH peak (CVF LH + 2 and CVF LH + 7, respectively). ESCs-EVs were isolated from both decidualized (dESCs-EVs) and not decidualized (eESCs-EVs) endometrial cells. Semen samples (n = 15) were collected from normozoospermic men. Participants/materials, setting, methods EVs were isolated using differential centrifugations and characterized using Transmission Electron Microscopy (TEM), Nanoparticle Tracking Analysis (NTA), and western blotting (WB). EVs capture by spermatozoa was assessed using flow cytometry. EVs impact on sperm progressive motility was evaluated following WHO guidelines. EVs potential to induce sperm capacitation was examined through sperm acrosome reaction (AR) induction using a Ca2+ ionophore, followed by Coomassie blue staining. All experiments were performed using EVs at 50 and 100 μg/mL. Main results and the role of chance Isolated EVs displayed comparable size, with mean diameters ranging from 147,9±11,6 nm to 168,1±6,2 nm (mean ± SEM) and average concentrations from 9,53E+10 ± 8,11E+09 to 2,70E+11 ± 3,49E+10 particles/ml (mean ± SEM). TEM analysis confirmed their round-shaped morphology and structural integrity, while WB confirmed their positivity for EVs markers (Alix, TSG101, CD63, CD9). Spermatozoa efficiency in capturing these fluorescently-labeled EVs, measured as the percentage of fluorescence-positive spermatozoa, reached up to 3,6±1,1% for FF-EVs, 12,1±3,7% for CVF-EVs and 9,4±1,9% for ESCs-EVs (mean ± SEM). Sperm cells co-incubated with any EVs group maintained unaffected vitality. Following a 4-hour incubation with EVs, a rise in the proportion of progressively motile spermatozoa was observed, with percentages of progressively motile spermatozoa ranging from a minimum of 45,4±3,8% for CVF-EVs LH + 7 to a maximum of 52,7±2,7% for dESCs-EVs (mean ± SEM). This increase resulted to be statistically significant (p < 0.05 and p < 0.01, respectively) compared to the untreated (UT) condition, which exhibited 26,2±0,7% of progressively motile sperm cells. Spermatozoa that completed the AR, indicating capacitation, after 4-hour incubation with EVs resulted to be up to 25,8±3,2% for FF-EVs, 35,0±6,6% for CVF-EVs and 26,9±3,0% for ESCs-EVs, compared to an average of 13% in the UT control. Limitations, reasons for caution This study constitutes a pilot investigation characterized by a limited sample size, necessitating larger cohorts to validate results. Wider implications of the findings Studied EVs may exert beneficial effects on capacitation and progressive motility also on asthenozoospermic (AS) men’s spermatozoa. Therefore, they could be used in ART cycles to prime spermatozoa from AS men and increase the percentage of sperm cells displaying progressive motility, potentially favouring fertilization via classical IVF instead of ICSI. Trial registration number not applicable

P-013 A potential therapeutic approach of intermittent fasting on sperm viscosity

Abstract Study question Does intermittent fasting affect sperm viscosity and motility? Summary answer The obtained results demonstrate that intermittent fasting significantly improves sperm viscosity and motility, and might be considered as a potential therapeutic approach for sperm hyperviscosity. What is known already Numerous factors influence sperm quality, functionality, and, consequently, male fertility. In the last decade, nutritional deficiency and lifestyle, have been grouped as popular factors that affect human sperm function and fertilization processes. Semen hyperviscosity which occurs in 12-29% of ejaculates is another condition that can contribute to male subfertility, as it is associated with changes in the chemical and physical characteristics of semen. Additionally, semen hyperviscosity contributes as well to poor sperm motility. Based on the aforementioned facts, focusing on identifying factors to enhance sperm viscosity and motility is crucial for improving IVF outcomes. Study design, size, duration The present study was based on semen analyses of 60 samples from 30 IVF patients. Following the purpose of the study and inclusion criteria, all 30 patients were diagnosed with abnormal sperm viscosity. Out of this number, 13 patients had asthenozoospermia, 4 patients were diagnosed with asthenoteratozoospermia, 7 patients were diagnosed with teratozoospermia and 6 patients had normozoospermia. All patients followed 4 days of sexual abstinence. All specimens were analyzed within 1 hour of collection. Participants/materials, setting, methods Sperm analysis including sperm viscosity was processed according to the WHO laboratory manual sperm criteria (Sixth edition 2021). For the study purpose, all the participants followed an intermittent fasting period of 14-16 hours for 30 days. Samples were analyzed twice, the first time before fasting and the second time after 30 days of fasting. Most of the patients did not consume food from 6 pm until 8 or 10 am. Main results and the role of chance The average age of the patients included in the study was 37.53 ± 4.6 years, while the BMI ranged from 21.26 to 32.28 kg/m2. Abnormal viscosity (hyperviscosity) was diagnosed when the drop formed a thread more than 2 cm long, while normal liquefied ejaculate fell as small discrete drops. The obtained results indicated that sperm parameters were improved after intermittent fasting. The most significant difference was observed in semen viscosity where out of the 30 patients who were diagnosed with abnormal semen viscosity after 30 days of intermittent fasting only 9 patients still had viscose semen (p<0.0001). Additionally, a significant difference was revealed in sperm motility before and after the fasting period as well(33.73±12.15 vs 44.10 ±11.54; respectively p < 0.001). Among the 17 subjects initially diagnosed with abnormal motility, following the implementation of intermittent fasting, 11 demonstrated a sperm improvement and were diagnosed with normal motility. The difference in sperm viability (52.8± 7.71 vs 61.3±8.37; respectively p < 0.01) and Ph values before and after fasting as well were statistically significant. On the other side, although sperm concentration (Mil/ml) was increased after fasting, the difference was not statistically significant (39.03±19.52 vs 41.53±19.19; respectively p = 0.92). Additionally, intermittent fasting did not affect sperm morphology either. Limitations, reasons for caution The limitation of the presented study was a relatively small patient cohort. Further research should link intermittent fasting with IVF outcomes. Wider implications of the findings Until today there are no clear dietary guidelines created for infertile males. As far as we know this is the first research, that includes a comparison of sperm parameters, based on intermittent fasting, and the obtained results confirm that intermittent fasting can be used as a potential therapeutic approach. Trial registration number not applicable

P-237 Fertilization prediction of mature oocyte based on microscopic image of first polar body morphology using YOLOv8

Abstract Study question Can a machine learning-based model predict successful fertilization of oocyte using first polar body morphology? Summary answer With small imbalance oocyte dataset, the most successful model to predict fertilization of mature oocyte is YOLOv8lcls with 97.47% accuracy, 100.00% specificity, and 94.87% sensitivity. What is known already The development of oocytes is crucial in the development and fertility of embryos in In Vitro Fertilization (IVF). There are several internal and external factors that could affect the fertilization competency of oocytes, one of which is the first polar body (PBI). The importance of the quality of PBI in ICSI has long been studied and has been evaluated by scoring systems regarding Istanbul Consensus. However, there is still a debate regarding the influence of PBI on the developmental competency of oocytes. Study design, size, duration This study involved the analysis of retrospective data including the microscopic light image of meiosis II (MII) oocytes at the time of Intracytoplasmic sperm injection (ICSI), along with polar body scoring by embryologists and the fertilization outcome of 1006 MII oocytes from a single public IVF clinic, between 2020-2022. After ICSI, fertilized oocytes were classified into 2 groups: 2PN and non-2PN (e.g., 0PN, 1PN, > =3PN). Participants/materials, setting, methods Light microscopic images of oocytes in the metaphase of meiosis II (MII) stage are captured, followed by the prediction of the likelihood of 2PN development after fertilization through intra-cytoplasmic sperm injection (ICSI). The image has its PBI segmented by the CNN and then cropped out of the original image. The PBI image is then fed into the prediction CNN to obtain the final result. The model performance was determined by evaluating accuracy, sensitivity, and specificity. Main results and the role of chance There were 1006 images of MII oocytes included in this study. After ICSI, 860 oocytes were fertilized resulting in the 2PN stage embryos and were classified as 2PN, whereas 146 oocytes were not fertilized or abnormally fertilized and were classified as non-2PN. Among these images, 782 oocytes received a PBI score of A, 677 of which are 2PN and 105 are non-2PN. The B grade consists of 224 images, of which are 183 2PN, and 41 non-2PN. The dataset was split into 3 subsets for training (70%), validation (10%), and testing (20%). The images are augmented by horizontal flipping to increase the amount of data and allow the model to observe several positions of the PBI. The YOLOv8 segmentation model is applied to segment the PBI and locate its position. The image is then rotated to move the PBI to the top and the PBI is cropped based on its position. Lastly, the cropped PBI image is used to predict the development by using the YOLOv8 Classify model trained on an augmented dataset. The YOLOv8l-cls model achieves the highest accuracy at 97.47%, specificity at 100.00%, and sensitivity at 94.87%. Limitations, reasons for caution The model does not include information on sperm quality, which also essentially contributes to the fertilization process. 2PN stage prediction is just the initial and surrogate outcome in IVF treatment. The model’s interpretation of the PBI score remains inconclusive. Therefore, additional experiments using dataset with balanced PBI grading are recommended. Wider implications of the findings The developed model is the first step in paving the way to establish and implement tools for assisting embryologists in selecting and prioritizing the high-potential oocyte for fertilization. Additional factors such as patient characteristics, other features of oocytes, and male factors should be further integrated to improve model performance. Trial registration number NA

P-504 Examining the success rates following a second embryo biopsy for inconclusive results in preimplantation genetic testing for monogenic diseases (PGT-M): a matched case-control study

Abstract Study question Can an embryo re-biopsy for a previous inconclusive result influence clinical pregnancy compared to a successful PGT-M diagnosis from the first biopsy? Summary answer The embryo re-biopsy intervention for PGT-M seems to reduce the implantation potential of a blastocyst. What is known already PGT-M allows embryo genetic profiling prior to transfer in couples with hereditary genetic disorders, as part of in vitro fertilization process. The need of a trophectoderm biopsy for the retrieval of a fragment of about 8-10 blastomeres is generally sufficient for optimal DNA amplification and molecular analysis. However, a proportion of embryos (approximately 10-12%) fails to be properly diagnosed after PGT-M testing due to inconclusive results. Therefore, as an attempt to obtain a genetic diagnosis, the embryo biopsy can be repeated on the same embryos although retesting involves a second round of biopsy and a second round of vitrification. Study design, size, duration Twenty-seven embryos from women undergoing PGT-M at an infertility center and transferred after two biopsies for genetic analysis were matched for age, fertility status, and study period, to embryos transferred after a conclusive PGT-M result after the first biopsy. The main outcome was the clinical pregnancy rate following embryo transfers in cases in whom embryos had a single biopsy, and when a re-biopsy was performed. The secondary outcome was the live birth rate. Participants/materials, setting, methods Ovarian stimulation, oocyte collection and fertilization were performed according to standard protocols. Embryos were cultured to blastocyst stage in groups of up to five, using one-step media. Blastocysts were biopsied on day 5, 6, or 7, employing either the pulling or flicking method based on their appearance. Biopsied cells were stored at -20 °C for genetic analysis. Vitrification was performed using Kitazato protocol. If results were inconclusive, a second biopsy was performed in surviving blastocysts. Main results and the role of chance The median [interquartile range] number of retrieved and injected oocytes was similar between the two groups, as well as fertilization rate and number of blastocysts obtained. No difference between the two groups was also observed pertaining to the day of embryo culture in which the first biopsy was performed. Conversely, the number of non-diagnosticated and transferable blastocysts was lower in patients whose blastocysts were re-biopsied. In 90% of the re-biopsies an informative result was obtained. Clinical pregnancy rate was 52% (95% CI: 34-69) following the transfer of a single-biopsy blastocyst and 30% (95% CI: 16-48) following the transfer of a re-biopsied blastocyst. The likelihood to have a healthy baby was 33% (95% CI: 19-52) following the transfer of a blastocyst biopsied once and 22% (95% CI: 11-41) following the transfer of a re-biopsied blastocyst. Limitations, reasons for caution The matching design, which aligns women’s age and fertility status in the two groups, allowed the assessment of pregnancy rates while avoiding potential confounding factors associated with retrospective analysis. Given that it is the first study with this design, a larger sample size is necessary to further elucidate these findings. Wider implications of the findings Despite the limited sample size, these results are of extreme clinical relevance. They offer valuable insights for guiding decisions about embryos with unclear results, contributing to the improvement of strategies for selecting those with the highest probability of pregnancy. Trial registration number not applicable

Long-Term In Vitro Culture Alters Gene Expression Pattern of Genes Involved in Ontological Groups Representing Cellular Processes.

The oviduct provides an optimal environment for the final preparation, transport, and survival of gametes, the fertilization process, and early embryonic development. Most of the studies on reproduction are based on in vitro cell culture models because of the cell's accessibility. It creates opportunities to explore the complexity of directly linked processes between cells. Previous studies showed a significant expression of genes responsible for cell differentiation, maturation, and development during long-term porcine oviduct epithelial cells (POECs) in vitro culture. This study aimed at establishing the transcriptomic profile and comprehensive characteristics of porcine oviduct epithelial cell in vitro cultures, to compare changes in gene expression over time and deliver information about the expression pattern of genes highlighted in specific GO groups. The oviduct cells were collected after 7, 15, and 30 days of in vitro cultivation. The transcriptomic profile of gene expression was compared to the control group (cells collected after the first day). The expression of COL1A2 and LOX was enhanced, while FGFBP1, SERPINB2, and OVGP1 were downregulated at all selected intervals of cell culture in comparison to the 24-h control (p-value < 0.05). Adding new detailed information to the reproductive biology field about the diversified transcriptome profile in POECs may create new future possibilities in infertility treatments, including assisted reproductive technique (ART) programmes, and may be a valuable tool to investigate the potential role of oviduct cells in post-ovulation events.